0576117 - ÚIACH 2024 IT eng A - Abstract
Foret, František - Voráčová, Ivona - Kocianová, Vanda - Hrušková, Helena
Epitachophoresis - high-efficiency method for concentration and separation of ionic compounds from large-volume samplesv.
ITP 2023. Abstracts. 2023. s. 37-37.
[International Symposium on Electro- and Liquid-Phase Separation Techniques /29./ ITP 2023. 13.09.2023-17.09.2023, San Felice Circeo]
Institutional support: RVO:68081715
Keywords : epitachophoresis * instrumentation * DNA * proteins
OECD category: Analytical chemistry

This communication reports on new instrumental systems for processing the crude samples by discontinuous electrophoresis in a circular arrangement – epitachophoresis (ETP) with almost unlimited concentration factor. Several experimental parameters have been studied, including the size, shape, and size of the zone stabilizing media and devices for large samples. Polyacrylamide or agarose gels are the most frequently used sieving and stabilizing media in slab gel electrophoresis; however, such sieving materials limit the size of the concentrated nucleic acids. In part of this work, we have also explored large pore materials and 3D printing to form rigid stabilizing manifolds to minimize liquid flow during the epitachophoresis. The device was printed using the stereolithography technique from a low water-absorbing resin. Different geometries of the 3D printed stabilizing manifolds were tested to concentrate ionic sample components in the anionic or cationic mode. Depending on its geometry, the devices can focus analytes from 1 to 50 ml of the sample into the collection cup with a size of 150 μL or less. Depending on the stabilization media and power used, the concentration time ranges from minutes to one hour. We have used the ETP to isolate DNA and RNA from biologically relevant samples in a single run, including formalin-fixed paraffin-embedded tissue. While the system was initially designed for extraction and focusing of nucleic acids, this presentation will also discuss the potential of the ETP for the separation and concentration of other analytes, including peptides and proteins.
Permanent Link: https://hdl.handle.net/11104/0345722